tbuckley/cabot-search huggingface dataset:
Abstract P1061: Salivary Extracellular Vesicle-associated LncRNAs As Novel Non-invasive Biomarkers For Acute Decompensated Heart Failure: The Pioneering SEAL-HF Study
Authors: Michail Spanos, Priyanka Gokulnath, Quanhu Sheng, Emeli Chatterjee, Guoping Li, Kathy Luong, Helge Immo Lehmann, Ashley Mingyi Wang, Chinemerem Nwokemodo-Ihejirika, Worawan Limpitikul, Dimitrios Varrias, David T Wong, Saumya Das
Journal: Circulation Research (2023)

Acute Decompensated Heart Failure (ADHF) is a major global health concern, and early diagnosis is critical for effective management. Non-invasive biomarkers have the potential to improve patient outcomes substantially. In this study, we investigated the presence of extracellular vesicle (EV)-associated long non-coding RNAs (lncRNAs) in plasma of ADHF patients and explored their non-invasive detection in saliva as the foundation of the SEAL-HF study. We performed RNA-sequencing on plasma-derived EVs from ADHF patients at admission and discharge and healthy controls to identify differentially expressed lncRNAs and mRNAs, using a fold change cutoff of ±1.5 and an adjusted p-value of &lt;0.05. Subsequent validation was carried out via qRT-PCR on plasma EV RNA samples from 182 patients. Salivary EVs were isolated and enriched, and their EV RNA content was profiled. Transcriptomic analysis unveiled several dysregulated EV-derived mRNAs and lncRNAs in ADHF patients compared to controls. Importantly, four lncRNAs (LINC00989, lnc-CALML5, AC092656.1, and RMRP) exhibited significant changes between the acute congestive phase and decongestion after diuresis and were found to circulate mainly within the EV compartment. Salivary EVs contained EV protein markers ALIX, CD81, and heart-derived protein marker Troponin T. qRT-PCR assay demonstrated the presence of LINC00989, lnc-CALML5, AC092656.1, and RMRP in salivary EVs, as well as cardiac-specific genes such as PLN. Our results highlight the potential of salivary EV-associated lncRNAs as non-invasive biomarkers for ADHF diagnosis, laying the groundwork for the SEAL-HF study and future clinical applications.
Abstract P1092: Endothelial Cell Aging Can Be Reversed By A Pharmacological, Partial Reprogramming Strategy
Authors: Katrin Kalies, Kai Knoepp, Leonie Wurmbrand, Jochen Dutzmann, Daniel Sedding
Journal: Circulation Research (2023)

Senescent endothelial cells (EC) are key players in the pathophysiology of cardiovascular diseases and characterized by a reduced angiogenic and regenerative potential. Therefore, reversing EC senescence represents a promising therapeutic strategy to restore vascular integrity and increase health and lifespan. Here, we show a reversal of EC senescence following the application of a pharmacological, non-genetic, partial reprogramming strategy to induce a timely restricted induction of the Yamanaka-factors Oct3/4, Sox2, Klf4 and c-Myc (OSKM). Methods to characterize the effects of pharmacological reprogramming included the quantification of gene expression as well as functional analysis of EC in vitro . In addition, the regenerative capacity of EC was evaluated in a hind-limb ischemia model in vivo in old C57BL/6J mice. We demonstrate that the application of a pharmacological cocktail of FDA approved drugs to senescent EC resulted in a robust but timely restricted induction of OSKM (p&lt;0.001). Additionally, siRNA-knockdown prior to the treatment confirmed the correlation between OSKM upregulation and pharmacological treatment. Induction of OSKM was associated with a significantly reduced expression of senescence markers as p16ink4a and p14arf (p&lt;0.001). Expression of cytokines as TNFa or Il-6 was significantly reduced compared to untreated senescent cells (p&lt;0.05). Further the telomere length was stabilized (p&lt;0.05). We also observed enhanced functional properties of senescent EC, such as proliferation, migration, sprouting and tube formation (p&lt;0.05). Long term cultivation of treated EC indicated that expression of p16ink4a and p14arf remained low (p&lt;0.05), and the migratory capacity was preserved. Non-senescent EC were not influenced by the pharmacological treatment. In vivo , a significantly improved blood flow was observed after hind limb ischemia in 22 months old C57BL/6J mice after 7 and 14 days (p&lt;0.001). Capillary density was significantly enhanced (p&lt;0.05). In conclusion, we demonstrate that a short induction of OSKM via a pharmacological approach holds the potential to reverse senescence in EC in vitro and thus to enhance endothelial regenerative capacity in vivo . This might present a promising strategy for ischemic diseases.
Abstract GS.12: A New Mouse Model To Investigate The Early Steps Of Arrhythmogenic Cardiomyopathy
Authors: Camilla Schinner, Lifen Xu, Ramón Castellanos-Martínez, Aude Zimmermann, Aude Zimmermann, Pauline Hanns, Gabriela M Kuster, Volker Spindler
Journal: Circulation Research (2023)

Arrhythmogenic Cardiomyopathy (ACM) is characterized by progressive loss of cardiomyocytes with fibrosis, systolic dysfunction and life-threatening arrhythmias. Mutations in genes of the desmosomal adhesion complex such as desmoglein-2 (Dsg2) are the major cause, but the underlying mechanisms leading to the disease are not well understood. Accordingly, only symptomatic treatment is available. Here, we establish an inducible Dsg2-W2A knock-in mouse model with temporal control of disease onset as new approach to analyse the early processes underlying the ACM phenotype. We have previously characterized a constitutive Dsg2-W2A knock-in model which develops a cardiac phenotype bearing all major criteria of ACM directly after birth. To control the onset of disease, we now generated a mouse line bearing the Dsg2-W2A mutation and a loxP -flanked Dsg2 on separate alleles, leading to functional hemizygosity of the Dsg2-W2A mutation upon tamoxifen-induced Cre expression. Detailed characterization of this iDsg2-W2A model revealed early repolarization changes in ECG recordings starting directly after induction and progression to extensive impairment of electrical conduction over 10 weeks. These changes were paralleled with the appearance of ventricular and supraventricular arrhythmias. Starting two weeks after induction, reduced cardiac output function of the right and later of the left ventricle was detectable via echocardiography. Finally, these mice developed structural changes with loss of cardiomyocytes and ventricular fibrosis similar to ACM patients. This study gives detailed insight on the early steps of ACM showing a sequence from changes in conduction, occurrence of arrhythmia to functional impairment, starting in the right and progressing to the left ventricle, together with structural changes of the heart. We expect this model to be valuable to discern patho-mechanisms in detail and to identify and test novel treatment options for ACM.
Abstract P3098: Immune Suppression Ameliorates A Mouse Model Of Lmna-related Cardiomyopathy With Nuclear Envelope Rupture
Authors: Atsuki En, Hanumakumar Bogireddi, Briana Thomas, Alexis Stutzman, Omar Almakki, Sachie Ikegami, Ivan P Moskowitz, Kohta Ikegami
Journal: Circulation Research (2023)

Lamin A/C (encoded by the LMNA gene) are the components of the nuclear lamina, a protein meshwork providing the structural integrity to the nuclear membrane. Dilated cardiomyopathy (DCM) caused by LNNA gene mutations is the second most common familial DCM in adults and associated with severe prognosis and high mortality. Despite decades of investigations into cell autonomous impacts of LMNA mutations, LMNA-DCM lacks effective treatments except for heart transplantation. Here, we identify cell non-autonomous effects of LMNA mutations and demonstrate the efficacy of targeting immune activities in mouse models of LMNA-DCM. We induced Lmna gene deletion specifically in cardiomyocytes in adult mice and tracked molecular, cellular, and physiological alterations in the heart at a high temporal resolution. We identified frequent localized nuclear envelope rupture, with the concomitant exposure of nuclear DNA to the cytoplasm, as the earliest direct consequence of Lamin A/C protein reduction in cardiomyocytes. Surprisingly, nuclear envelope rupture did accompany cardiomyocyte cell death or extensive transcriptional changes or cytosolic DNA sensing pathway activation within the cardiomyocytes. Instead, it coincided with extensive proinflammatory signaling activation within the cardiac macrophage population. This was followed by extensive fibrosis and structural and functional alterations of the heart, culminating in DCM and death. Treatment of LMNA-DCM mice with the immunosuppressant Dexamethasone suppressed fibrosis, profoundly improved the cardiac structure and function, and conferred a two-fold lifespan extension after Lmna gene deletion. This work suggested that immune activation is the major pathological component of LMNA-DCM and an effective target for treating LMNA-DCM patients.
Abstract P3059: Recombinant Adeno-associated Viral Vector 2/8 Model Of Interleukin 6 Mediated Chronic Inflammation
Authors: Johanna E Jacobs, Wannes Maes, Ellen Caluwe, Marleen Lox, Hilde Gillijns, Peter Pokreisz, Tania Roskams, Chris Van Den Haute, Rik Gijsbers, Stefan P Janssens, Lucas Van Aelst
Journal: Circulation Research (2023)

Background: Interleukin 6 (IL6) is a pleiotropic cytokine mostly known as a proinflammatory cytokine. There is a need for a reliable in vivo model to study IL6 mediated chronic inflammation. Hypothesis: The aim of our study was to create a mouse model mimicking the chronic inflammatory state as observed in cancer and inflammatory diseases. We opted for a recombinant adeno-associated viral vector (rAAV) to mimic the acute initiation of chronic inflammation later in life. Methods: We opted for a rAAV2/8 subtype, human EF1a short (EFS) promoter and firefly luciferase led by an internal ribosome entry site (IL6-IRES-fLuc) DNA cassette to create a stable low-grade expression of IL6 in liver and serum, with minimal expression in other organs, particularly the heart. rAAV2/8_EFS-IL6-IRES-fLuc and the control vector rAAV2/8_EFS-MCS-IRES-fLuc were generated and different dosages were tested. Results: All mice treated with high dose AAV2/8-IL6 vector (8.8* 10 10 genome copies (GC)/animal) showed severe weight loss and hepatosplenomegaly, starting between week 4-6 post injection. Histological examination of the liver showed features of chronic hepatitis. Infiltrating cells were found in both the liver and the heart. In the spleen, the histological structure of red and white pulp was disturbed and more giant cells could be detected. Cardiac function remained normal at 6 weeks post injection. Assuming that the previous results were due to an overexpression of IL6 due to too high amounts of rAAV2/8_EFS-IL6-IRES-fLuc, we subsequently challenged mice with a rAAV2/8_EFS-IL6-IRES-fLuc dilution series to test which vector dose could be used for reliable long-term low grade IL6 overexpression. After extensive testing, the ideal dose for a 12 weeks IL6 overexpression model was between 2.2 and 2.93* 10 10 GC. Using this dose range, we saw stable overexpression of IL6 without weight loss. Hepatosplenomegaly, disturbed anatomy of the spleen and infiltrating cells in the liver were still present yet less pronounced. Conclusion: We successfully generated and characterized a mouse model of IL6 overexpression to mimic chronic IL6 mediated inflammation. To the best of our knowledge this is the first model employing IL6 overexpression using a rAAV viral vector.
Abstract P3112: The Effects Of Sex Hormones On Intestinal Lipoprotein Size
Authors: Andromeda M Nauli, Ann Phan
Journal: Circulation Research (2023)

Larger intestinal lipoproteins are more likely to be retained in the intestinal wall, allowing their fat to be hydrolyzed and taken up by the abdominal viscera. Since men generally accumulate more abdominal visceral fat than women, we hypothesize that testosterone stimulates the release of larger intestinal lipoproteins relative to estrogen and progesterone. To test our hypothesis, the differentiated Caco-2 cells (n ≥ 12 per group) were induced to produce intestinal lipoproteins while simultaneously subjected to the physiological concentrations of sex hormones by using our semipermeable membrane system. The lipoproteins were isolated by using density gradient ultracentrifugation, and the amount of ApoB and TG were determined. Since each lipoprotein consists of only one ApoB, higher ratios of TG/ApoB suggest larger lipoproteins as shown by our previous studies. Our results indicated that while there was no dose-dependent effect of beta-estradiol (p &gt; 0.05) and progesterone (p &gt; 0.05), testosterone significantly (p = 0.004) increased the TG/ApoB ratio. When these hormones were combined to resemble the plasma concentrations observed in men and the different phases of the ovarian cycle (follicular, ovulatory, and luteal), both the male (p = 0.045) and the luteal phase (p = 0.043) had significantly higher TG/ApoB ratios than the ovulatory phase. The ovulatory phase secreted significantly less amount of TG than the other groups, and the follicular phase secreted significantly less amount of TG than the male (p = 0.0003). ApoB was comparable among all these four groups (p &gt; 0.05). In conclusion, unlike beta-estradiol and progesterone, testosterone significantly increased the size of intestinal lipoproteins. The ovulatory phase secreted the least amount of TG and produced significantly smaller intestinal lipoproteins than the male and the luteal phase. These studies support our hypothesis that through their testosterone effects, men produce larger intestinal lipoproteins. Larger lipoproteins tend to stay longer in the intestinal wall, allowing their fat to be taken up preferentially by the abdominal viscera. Our studies may partly explain why men are more likely to accumulate abdominal visceral fat, which is an independent predictor of mortality.
Abstract P3168: Identification Of Exosome MicroRNA In Association With Severe Calcification In Coronary Chronic Total Occlusion
Authors: Ji-Hye Son, Hun Soo Chang, Inki Moon, Min Gyu Kong, Su Yeong Park, Dongeon Kim, Hyung Oh Choi, Hyun-Woo Park, Yoon Haeng Cho, Heon Lee, Jon Suh
Journal: Circulation Research (2023)

Backgrounds: Coronary artery calcification (CAC) is prevalent in patients with coronary chronic total occlusion (CTO). We investigated the association between plasma miRNAs and coronary artery calcification in patients with CTO. Methods: Blood samples were collected from 18 patients who underwent percutaneous coronary intervention (PCI) of CTO. Exosomal small RNA sequencing was performed from plasma to quantify miRNA expression. Calcium volume was quantified in computed tomography (CT) in CTO. Gene ontology and functional pathway were analyzed in the miRNAs that significantly correlates with calcium volume. Results: Calcium volume was positively correlated with miR-1-3p and miR-708-3p (p &lt;0.05), and was negatively correlated with 4 miRNAs -including let-7d-5p (p &lt;0.05). Gene enrichment and pathway analysis showed that the target genes of the 2 positively correlated miRNAs were involved in regulation of osteoblast differentiation and osteopontin signaling pathway. In contrast, the target genes of the 4 negatively correlated miRNAs were enriched in acetylcholine-mediated vasodilation involved in regulation of systemic arterial blood pressure, blood vessel development and response to sterol. Multiple linear regression showed that miR-1-3p and let-7d-5p could be key predictors of the calcium volume of CTO (adjusted R 2 = 0.798 and p &lt;0.001). Conclusions: We identified miR-1-3p and let-7d-5p could be related with CAC in patients with CTO.
Abstract P3043: Mir92b-3p: A Key Regulator In Vascular Homeostasis And Remodelling
Authors: Jan Michael Köster, Katrin Kalies, Solveig Boettcher, Fiene Daniel, Laura Hehl, Simon Guerlach, Kai Knoepp, Jochen Dutzmann, Daniel SEDDING
Journal: Circulation Research (2023)

In vascular disease, cells undergo important alterations in cellular function, influencing vascular homeostasis. Apoptosis is one of these functions, affecting vascular tone and plaque stability. Influencing this process for example, by targeting microRNAs is key for therapeutic strategies tackling vascular diseases in a cell-specific manner. We identified the critical role of miR92b-3p on the functional properties of vascular cells. Studying human primary coronary artery smooth muscle cells (SMCs) and human umbilical vein endothelial cells (ECs), methods included expression analysis in mice tissue after restenosis by wire-induced injury and in atherosclerosis. The role of miR-92b on cellular function and target regulation was determined by pre- and antagomir transfection. Initial analysis in vivo showed miR92b-3p to be significantly upregulated in atherosclerosis (p&lt;0,01) and regulated in restenosis (p&lt;0,05). Under growth conditions in both ECs and SMCs, miR-92b-3p was significantly higher expressed (p&lt;0,05). In SMCs, upregulation of miR92b-3p resulted in increased proliferation (p&lt;0,01) and significantly enhanced cell migration in SMCs (p&lt;0,001) but hampered migration in ECs (p&lt;0,01). Downregulation of miR-92b-3p in SMC resulted in decreased migrational (p&lt;0.001) and proliferative capacity (p&lt;0.001), accompanied by a reduction in metabolic activity (p&lt;0.05) and, exclusively in SMCs, increased apoptosis (p&lt;0.05). In ECs, proliferation was enhanced following downregulation (p&lt;0,01). Multiple targets were identified, using Western blot and qPCR including KLF4 (p&lt;0,01), DKK3 (p&lt;0,05), Notch1 (p&lt;0,01), PTEN (p&lt;0,05), and especially in SMC, Bim (p&lt;0,001). Regulation of Bim by miR92b-3p and subsequent apoptosis was then evaluated by siRNA-knockdown experiments, confirming their strong interaction. We further verified the crucial role of miR-92b-3p by reversing cell death in apoptotic SMCs through miR-92b-3p overexpression experiments (p&lt;0.05). This study reports miR 92b-3p to be a crucial regulator of vascular function. Therapeutic regulation might present a promising strategy for stabilizing critical plaques, restoring the function of vascular cells, and thereby preventing vascular remodeling.
Abstract P2081: Defective Translation Of Mitochondrial Proteins Impairs Cardiac Metabolism In HFpEF
Authors: Navid Koleini, Mariam Meddeb, Edwin Yoo, James Saba, Mohammad Keykhaei, Abigail Mulligan, Myo Htet, Farnaz Farshidfar, Raihan Kabir, Seoyoung Kwon, Olivia Hanselman, Desirae McKoy, Liang Zhao, Virginia S Hahn, Kavita Sharma, Simone Sidoli, Rachel Green, Mark J Ranek, Erika Pearce, David A Kass
Journal: Circulation Research (2023)

We recently reported that heart failure with preserved ejection fraction (HFpEF) in human myocardium has broadly reduced acylcarnitines and Krebs cycle intermediates, and that transcriptomics shows increased gene expression of oxidative-phosphorylation (OxPhos) related pathways. However, new preliminary data finds protein expression related to OxPhos is decreased. Here, we explored a key mechanism that could drive depressed mitochondrial protein translation in HFpEF. Elongation factor 5A (EIF5A) is a ribosomal subunit that is post-translationally modified on lysine 50 by the transfer of an aminobutyryl moiety from spermidine, called hypusination (Hyp). We find EIF5A Hyp and the mitochondrial proteome are diminished despite elevated gene transcripts in the myocardium of human and a mouse model of HFpEF (latter high-fat diet +L-NAME). Inhibiting EIF5A hypusination in neonatal rat cardiomyocytes results in ribosome stalling, disruption of OxPhos, and increased glycolysis.Proteomics of adult cardiomyocytes with inhibition of the rate-limiting hypusination enzyme deoxyhypusine synthase (DHPS) phenocopies depressed mitochondria as in human HFpEF. Cardiomyocyte-specific knockout of DHPS is lethal in pups within 24hrs after birth. These mice have significantly less myocardial EIF5A Hyp . Cardiac-specific DHPS+/- are viable and have no heart phenotype in unstressed situations, but display marked cardiac dilation with low ejection fraction with failure to hypertrophy in response to transverse aortic constriction.Four weeks following tamoxifen-inducible homozygous knockout of DHPS in cardiomyocytes of 8-week-old mice, we find impaired diastolic function (increased E/E’) with preserved ejection fraction and reduced exercise tolerance. Despite reduced hypusination in HFpEF, ornithine levels are elevated suggesting inhibition of hypusination at the hypusinating enzymes. Interestingly, DHPS levels are decreased in human HFpEF hearts.In summary, we have uncovered a novel role of polyamines in cardiomyocytes: the facilitation of translation of mitochondrial proteins by EIF5A hypusination. We find reduced hypusination resulting from defects in hypusine synthesizing enzymes contributes to mitochondrial defects in HFpEF.
Abstract P3086: Aldose Reductase Deletion Protects Ischemic Murine Hearts By Modulating Immune Cell Dynamics
Authors: Syed Nurul Hasan, Gautham Yepuri, Nosirudeen Quadri, Sangeeta Bhallamudi, Rollanda Bernadin, Kaamashri Mangar, Ann Schmidt, Ravichandran Ramasamy
Journal: Circulation Research (2023)

Cardiac injury remains a major cause of death worldwide. Previous studies have demonstrated that chronic unchecked inflammation, during post ischemic reperfusion, adversely affects adaptive repair and augments tissue damage. Nature of the infiltrating immune cells determines the extent of repair vs exacerbation of tissue damage during post myocardial ischemia. Understanding mechanisms by which ischemia-mediated inflammatory responses exacerbate tissue damage and mitigate repair pathways are vital to discover therapeutics for myocardial infarction. Our earlier studies demonstrated that aldose reductase (AR) is a key mediator of myocardial ischemia-reperfusion (I/R) injury. Specifically, it established that cardio myocyte AR drives metabolic and signalling mechanisms leads to injury and impaired function in hearts. In this study we investigated the role of AR in influencing time dependent changes in recruitment of immune cells, during in vivo myocardial I/R injury. AR knockout (ARKO) and littermate controls (WT) mice were subjected to LAD sugery followed by reperfusion ( In vivo I/R) and the temporal changes in recruitment of immune cells were determined at earlier (day 3) and later (day 7) time points. We found ablation of AR leads to significant (p&lt;0.0001) cardiac protection during post I/R in mice. Cardiac function measurements revealed greater recovery of ejection fraction and fractional shortening after I/R in ARKO vs WT hearts (p&lt;0.05). Deletion of AR leads to a significant (p =0.0053) reduction in T cells infiltration post I/R in the heart. We observed 2-fold reduction in CD8+ cells, and 2-fold increase in CD4+ cells in ARKO vs WT mice hearts post I/R injury. Specifically, 2-fold reduction in pro-inflammatory Th1, Th2 and Th17 cells and 3-fold increases in anti-inflammatory Treg cells (p = 0.0091) were observed in ARKO vs WT hearts at post I/R injury. In addition, we observed reduction in infiltration of pro-inflammatory macrophages, dendritic cells, and B cells in ARKO vs WT hearts post I/R. Taken together, our data suggests that deletion of AR protects I/R hearts by promoting increases in anti-inflammatory immune cells and paves the way for addressing immune cell-cardiomyocyte crosstalk mechanisms by which AR deletion protects from I/R injury.